Alcohol-Induced Alternative Splicing in Drosophila Memory Circuits

果蝇记忆电路中酒精诱导的选择性剪接

基本信息

批准号：
10677574
负责人：
Tariq Brown
金额：
$ 4.77万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677574
关键词：
Adult Affect Alcohol consumption Alcohols Alternative Splicing Amino Acids Axon Behavior Behavior assessment Behavioral Biological Assay Brain Calcium Cells Cellular Neurobiology Communication Corpus striatum structure Cues Custom Data Analyses Dendrites Development Dopamine Dopamine D2 Receptor Drosophila genus Drosophila melanogaster Environment Ethanol Event Excision Experimental Designs Fc Receptor Fellowship Future Gene Expression Genes Genetic Genome Genome engineering Goals Head Human Image In Vitro Individual Institution Intoxication Locomotion Mammals Measures Memory Mentors Modeling Molecular Mushroom Bodies Nervous System Physiology Neurons Neurosciences Odors Oral Output Pathology Pattern Phenotype Positioning Attribute Post-Translational Modification Site Post-Translational Protein Processing Property Protein Isoforms Proteins RNA Processing RNA Splicing Relapse Retrieval Rewards Rodent Role Sensory Serine Sucrose Technical Expertise Testing Tissues Training Transcript Universities Variant Ventral Tegmental Area Work Writing alcohol exposure alcohol memory alcohol response alcohol use disorder behavior change behavior influence career cell type chronic alcohol ingestion dopaminergic neuron experience experimental study fly genetic approach genetic manipulation knock-down memory encoding memory retrieval mutant neural neural circuit neurogenetics neuropsychiatric disorder postsynaptic preference presynaptic prevent programs protein protein interaction receptor receptor function response sugar superresolution imaging temporal measurement two-photon

项目摘要

Repeated alcohol experiences can produce long-lasting memories for sensory cues associated with intoxication. These memories can problematically trigger relapse in individuals recovering from alcohol use disorder (AUD). The molecular mechanisms by which ethanol changes memories to become long-lasting and inflexible remain unclear. We recently demonstrated that formation of these memories results in expression of alternative transcript isoforms in memory-encoding neurons in Drosophila melanogaster. Drosophila rely on mushroom body (MB) neurons to make associative memories, including memories of ethanol-associated sensory cues. Decreasing expression of genes that play a role in splicing in adult MB neurons reduces formation of these memories, demonstrating the necessity of RNA processing in ethanol memory formation. Moreover, decreasing expression of genes that are alternatively spliced like the Dopamine-2-like Receptor (Dop2R) in adult MB neurons reduces ethanol memory formation. This suggests that the splicing changes in these genes has functional implications for future memory formation. The central hypothesis of this proposal is that the dynamic alternative splicing of Dop2R in response to alcohol exposure directs reward behavior by changing the activity of neural circuits. We generated mutant Drosophila that have forced expression of the naïve or trained Dop2R isoform, which allows us to interrogate how these splice variants affect neural dynamics and behavior at a highly mechanistic level. The applicant will reveal intracellular localization of DOP2R protein (Aim 1, how alternative splicing of Dop2R affects reward memory (Aim 2), and plasticity within memory circuits (Aim 3). By completing these aims, the applicant will gain scientific and technical expertise in cellular neurobiology and neurogenetics, behavioral assessment of Drosophila, super-resolution imaging, and 2-photon calcium imaging. Through a comprehensive training plan, this fellowship will facilitate the professional development of the applicant in robust experimental design and data analysis; written and oral scientific communication; and effective and inclusive mentoring. Successful completion of the project and training goals are fully supported by the interactive and supportive institutional environment of Brown University and in the Neuroscience Graduate Program, and will prepare the applicant for the next steps towards an independent scientific career.

重复的饮酒经历可以对与中毒相关的感官线索产生持久的记忆。这些记忆可能会引发酒精使用障碍 (AUD) 康复者的旧病复发。乙醇使记忆变得持久且不灵活的分子机制仍然存在我们最近证明这些记忆的形成会导致替代性的表达。果蝇记忆编码神经元中的转录亚型依赖于蘑菇体。 (MB) 神经元产生联想记忆，包括与乙醇相关的感觉线索的记忆。减少在成年 MB 神经元剪接中发挥作用的基因的表达会减少这些基因的形成记忆，证明了 RNA 加工在乙醇记忆形成中的必要性。成人 MB 中选择性剪接的基因的表达，如多巴胺 2 样受体 (Dop2R) 这表明这些基因的剪接变化减少了乙醇记忆的形成。该提案的中心假设是动态的。 Dop2R 响应酒精暴露的选择性剪接通过改变活性来指导奖励行为的神经回路。我们生成了突变型果蝇，它强制表达原始或经过训练的 Dop2R 同工型，该亚型使我们能够探究这些剪接变体如何以高度机械的方式影响神经动力学和行为申请人将揭示DOP2R蛋白的细胞内定位（目标1，如何选择性剪接） Dop2R 影响奖励记忆（目标 2）和记忆回路内的可塑性（目标 3）。申请人将获得细胞神经生物学和神经遗传学、行为学方面的科学和技术专业知识通过全面评估果蝇、超分辨率成像和 2 光子钙成像。培训计划，该奖学金将促进申请人在强大的实验方面的专业发展设计和数据分析；书面和口头科学交流；以及有效和包容性的指导。项目和培训目标的成功完成得到了互动和支持的充分支持布朗大学的制度环境和神经科学研究生项目，并将准备申请者走向独立科学事业的下一步。